Skip to main content
NCBI home page
Applied Microbiology logoLink to Applied Microbiology
. 1965 Sep;13(5):706–712. doi: 10.1128/am.13.5.706-712.1965

Steroid Lysis of Protoplasts and Effects of Stabilizers and Steroid Antagonists

Rodney F Smith 1,1, Donald E Shay 1
PMCID: PMC1058329  PMID: 4956242

Abstract

Six synthetic antimicrobial steroids were examined for indications of their mechanism of action. Dequadin acetate, cetyl pyridinium chloride (CPC), and sodium deoxycholate were studied for comparison. Aerated cells of Sarcina lutea were washed, suspended in 1.06 M sucrose, and converted to protoplasts with 20 μg/ml of lysozyme. Lysis was measured optically at 650 mμ as a decrease in optical density. Screening tests with 50 μg/ml of each compound showed five steroids and CPC to be lytic. Protoplasts were strongly protected from lysis by pretreatment with 0.001 to 0.004 M spermine tetrahydrochloride. Other polyamines, such as spermidine phosphate, were less protective, and putrescine was ineffective. Uranyl nitrate (5 × 10-4 M) rapidly agglutinated protoplasts and protected them from rupture by the lytic agents. Similar studies with 0.001 to 0.004 M Mg++ showed varying degrees of protection, which, in most cases, was only temporary. Steroidal lysis did not appear to be related to chelation, since ethylenediaminetetraacetate did not cause lysis alone and antagonized some lytic compounds. Lecithin, Tween 80, Tween 20, and Span 20 at 0.05% exhibited certain effects on protoplast stability. Span 20 strongly prevented lysis by steroids. Tween 20 alone quickly caused protoplast rupture. Lecithin and Tween 80, which also caused lysis alone, interfered with lytic steroids and CPC. The test compounds were both inhibitory and lethal to cells of Sarcina lutea. The results suggest that direct action on cell membranes may be chiefly responsible for the antimicrobial properties of the steroids.

Full text

PDF
706

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BROCK T. D. Effects of magnesium ion deficiency on Escherichia coli and possible relation to the mode of action of novobiocin. J Bacteriol. 1962 Oct;84:679–682. doi: 10.1128/jb.84.4.679-682.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BROWN A. D. ASPECTS OF BACTERIAL RESPONSE TO THE IONIC ENVIRONMENT. Bacteriol Rev. 1964 Sep;28:296–329. doi: 10.1128/br.28.3.296-329.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. GALE G. R. CYTOLOGY OF CANDIDA ALBICANS AS INFLUENCED BY DRUGS ACTING ON THE CYTOPLASMIC MEMBRANE. J Bacteriol. 1963 Jul;86:151–157. doi: 10.1128/jb.86.1.151-157.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. GALE G. R., WELCH M., HYNES J. B. Pharmacology of certain antimicrobial N1, N5-substituted biguanides. J Pharmacol Exp Ther. 1962 Nov;138:277–284. [PubMed] [Google Scholar]
  5. GILBY A. R., FEW A. V. Lysis of protoplasts of Micrococcus lysodeikticus by ionic detergents. J Gen Microbiol. 1960 Aug;23:19–26. doi: 10.1099/00221287-23-1-19. [DOI] [PubMed] [Google Scholar]
  6. HAROLD F. M. STABILIZATION OF STREPTOCOCCUS FAECALIS PROTOPLASTS BY SPERMINE. J Bacteriol. 1964 Nov;88:1416–1420. doi: 10.1128/jb.88.5.1416-1420.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hancock R., Fitz-James P. C. Some differences in the action of penicillin, bacitracin, and vancomycin on Bacillus megaterium. J Bacteriol. 1964 May;87(5):1044–1050. doi: 10.1128/jb.87.5.1044-1050.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. NEWTON B. A. Site of action of polymyxin on Pseudomonas aeruginosa: antagonism by cations. J Gen Microbiol. 1954 Jun;10(3):491–499. doi: 10.1099/00221287-10-3-491. [DOI] [PubMed] [Google Scholar]
  9. RAZIN S., ROZANSKY R. Mechanism of the antibacterial action of spermine. Arch Biochem Biophys. 1959 Mar;81(1):36–54. doi: 10.1016/0003-9861(59)90173-0. [DOI] [PubMed] [Google Scholar]
  10. ROGERS D. SOLUBILIZATION OF PROTEIN ACCOMPANYING LOSS OF PERMEABILITY OF ESCHERICHIA COLI. J Bacteriol. 1964 Aug;88:279–292. doi: 10.1128/jb.88.2.279-292.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. ROODYN D. B. Protein synthesis in mitochondria. 3. The controlled disruption and subfractionation of mitochondria labelled in vitro with radioactive valine. Biochem J. 1962 Oct;85:177–189. doi: 10.1042/bj0850177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. SHOCKMAN G. D., LAMPEN J. O. Inhibition by antibiotics of the growth of bacterial and yeast protoplasts. J Bacteriol. 1962 Sep;84:508–512. doi: 10.1128/jb.84.3.508-512.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. SMITH R. F., SHAY D. E., DOORENBOS N. J. ANTIMICROBIAL ACTION OF NITROGEN-CONTAINING STEROIDS. J Bacteriol. 1963 Jun;85:1295–1299. doi: 10.1128/jb.85.6.1295-1299.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SMITH R. F., SHAY D. E., DOORENBOS N. J. EFFECTS OF PROTEIN, LIPIDS, AND SURFACTANTS ON THE ANTIMICROBIAL ACTIVITY OF SYNTHETIC STEROIDS. Appl Microbiol. 1963 Nov;11:542–544. doi: 10.1128/am.11.6.542-544.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SMITH R. F., SHAY D. E., DOORENBOS N. J. RELATIONSHIP OF SURFACTANT PROPERTIES OF SOME SYNTHETIC STEROIDS TO BACTERICIDAL ACTION. J Pharm Sci. 1964 Oct;53:1214–1216. doi: 10.1002/jps.2600531019. [DOI] [PubMed] [Google Scholar]
  16. SOCRANSKY S. S., LOESCHE W. J., HUBERSAK C., MACDONALD J. B. DEPENDENCY OF TREPONEMA MICRODENTIUM ON OTHER ORAL ORGANISMS FOR ISOBUTYRATE, POLYAMINES, AND A CONTROLLED OXIDATION-REDUCTION POTENTIAL. J Bacteriol. 1964 Jul;88:200–209. doi: 10.1128/jb.88.1.200-209.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. STALHEIM O. H., WILSON J. B. CULTIVATION OF LEPTOSPIRAE. II. GROWTH AND LYSIS IN SYNTHETIC MEDIUM. J Bacteriol. 1964 Jul;88:55–59. doi: 10.1128/jb.88.1.55-59.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tabor C. W. STABILIZATION OF PROTOPLASTS AND SPHEROPLASTS BY SPERMINE AND OTHER POLYAMINES. J Bacteriol. 1962 May;83(5):1101–1111. doi: 10.1128/jb.83.5.1101-1111.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES